KR101385644B1 - A cosmetic composition for preventing hair loss containing an extract of weigela subsessilis by bioconversion - Google Patents

Abstract

The present invention relates to a cosmetic composition to prevent hair loss and promote hair growth which includes extract of fermented Weigela subsessilis which is bioconverted into Ganoderma lucidum mycelium and, more specifically, to a cosmetic composition to prevent hair loss and promoting hair growth which includes filtrate as an active ingredient which is obtained by the following steps: extracting extract from the Weigela subsessilis; culturing after mixing the Ganoderma lucidum mycelium or culture medium of the Ganoderma lucidum mycelium with the extract; and removing the Ganoderma lucidum mycelium after culturing. The present invention promotes hair growth without stimulating scalp, promotes activities of hair root cells, and promotes hair growth through antiflammation effects for alopecia caused by inflammation. [Reference numerals] (AA) First week; (BB) Third week; (CC) Example 1; (DD) Positive control group (3% minoxidil); (EE) Negative control group

Description

A cosmetic composition for preventing hair loss and promoting hair growth containing fermented fern extract {A COSMETIC COMPOSITION FOR PREVENTING HAIR LOSS CONTAINING AN EXTRACT OF WEIGELA SUBSESSILIS BY BIOCONVERSION}

The present invention is a bioconverted birches ( Weigela subsessilis ) relates to a hair loss preventing and hair growth cosmetic composition containing an extract, and more particularly, to a hair loss preventing and hair growth promoting cosmetic composition containing the filtrate obtained by fermenting the extract of the young flower as an active ingredient.

Hair is a solid cylindrical fiber that only mammals have and is made of hardened, keratinized epithelial cells. Hair grows under normal conditions, stops growing for a while, then falls out and is replaced. In other words, the period of anagen, catagen, and telogen are repeated periodically. Hair growth of the scalp in adults is about 3 years, degenerative period is 3 weeks, and resting period is about 3 months. This is called the hair cycle, and since human hair has its own hair cycle, it always maintains a constant hair count without molting.

In general, the causes of hair loss include genetic factors, progression of aging, drug use, radiation therapy and post-stress stress.The main causes of hair loss from a biochemical and physiological point of view are hyperactivity, problems with scalp blood circulation and hair. It is known to lack nutrients essential for metabolism. Although these factors alone can cause hair loss, a combination of hair loss factors may lead to synergism, accelerating alopecia and worsening symptoms.

In recent years, as the stress index of modern people increases, hair loss symptoms occur not only in middle-aged and middle-aged men but also young men in their 20s and 30s, and hair and scalp injuries due to shampoo, mousse, perm, dyeing, and dry heat are caused. It has been shown to cause large hair loss.

Various oral and external medicines are used. The oral drug, finasteride, which is approved by the US FDA, inhibits the production of 5α-dihydrotestosterone, a testosterone that causes hair loss, and 6 months to 1 month. It is known that hair loss suppression or hair growth effects occur when taken for about a year, but when it is stopped, there is a problem of returning to its original state within two months, and sexual dysfunction such as decreased libido and erectile dysfunction and women have side effects such as risk of birth defects. Minoxidil, approved by the US FDA as an external preparation, has capillary expansion to prevent hair loss by applying it to the scalp, but it must be applied twice a day continuously and the drug stays on the scalp for 3 to 4 hours to have some effect. There is discomfort that appears. In addition, the corresponding effects are insignificant, and side effects such as itching and irritation appear.

Therefore, there is an urgent need for the development of a natural hair loss preventing agent that can be widely used by the general public without any side effects.

Therefore, the present inventors have studied the method to further increase the efficacy of the antioxidant effect of the locust extract among the plants that grow in nature, as a result of Ganoderma mushroom ( Ganoderma) lucidum ) When bioconversion to mycelium confirmed that the antioxidant effect, antimicrobial effect and anti-inflammatory effect and hair growth promoting effect significantly increased and completed the present invention.

Therefore, the present invention is to provide a cosmetic composition for preventing hair loss and promoting hair growth, comprising a filtrate obtained by bioconverting the extract of Weigela subsessilis into Ganoderma lucidum mycelium as an active ingredient. The purpose.

In order to achieve the above object, the present invention comprises the steps of extracting the extract from the bottle flower; Mixing the whole culture solution of Ganoderma lucidum mycelium or Ganoderma lucidum mycelium to the extract, followed by main culture; Removing the Ganoderma lucidum mycelium after the present culture; provides a cosmetic composition for preventing hair loss and promoting hair growth, comprising the filtrate obtained from the active ingredient.

Hereinafter will be described in detail with respect to the hair loss prevention and hair growth cosmetic composition containing the fermented young flower extract of the present invention.

In the case of extract extracted simply from the sol-tree as a solvent shows an effective efficacy in vitro, when applied to the skin cosmetics there is a problem that the effect is not substantially satisfactory.

On the other hand, the present inventors confirmed that the main component that shows the efficacy effect in the bottle extract of the flavonoids. Plant-rich phenolic compounds such as flavonoids, tannins, and anthocyanins are excellent antioxidant candidates for protecting skin damaged by UV rays. In particular, flavonoids or derivatives thereof have radical scavenging ability and lipid peroxidation. It is reported that antioxidant activity, such as a production inhibitory ability, is excellent.

In general, hydrophobic substances are more effective for skin penetration than hydrophilic substances. The ceramide component distributed in the stratum corneum is hydrophobic, which is more effective for interacting with hydrophobic substances than hydrophilic substances, so that the hydrophobic substance is more easily skinned. Because it can pass through.

Flavonoids, on the other hand, are mostly glycosides in which sugars are bound to one or more molecules, and are present in nature, have a relatively high molecular weight, and have some water-soluble properties. There is a problem that is difficult.

Therefore, the present invention comprises the steps of extracting the extract from the medicinal tree to provide a cosmetic composition for preventing hair loss and promoting hair growth that is easy to penetrate the skin as an effective skin external preparation; Mixing the whole culture solution of Ganoderma lucidum mycelium or Ganoderma lucidum mycelium to the extract, followed by main culture; Removing the Ganoderma lucidum mycelium after the main culture; containing the filtrate obtained from the above step as an active ingredient, the filtrate obtained through the above step has a total phenol content and total flavonoid content is increased than the extract extracted from the young The inventors confirmed that.

In addition, the present invention is the filtrate contained as an active ingredient is excellent in the antioxidant effect, antibacterial effect, promotes hair growth and hair root cell activity, less irritation to the skin, excellent anti-inflammatory effect through the anti-inflammatory action It is characterized by.

In addition, the present invention relates to a cosmetic composition for preventing hair loss and promoting hair growth containing a fermented vine extract, characterized in that the content of the extract is 0.001 to 30.0% by weight based on the total skin cosmetic composition. When the extract is included in less than 0.001% by weight, there is almost no hair growth effect, and when the extract is more than 30.0% by weight, the effect of increasing the effect of the increase in content is insignificant and economical.

In addition, the present invention relates to a cosmetic composition, characterized in that using the aqueous solution containing 40 to 95% (v / v) of ethanol as the extraction solvent during the extraction.

In addition, the present invention relates to a cosmetic composition, characterized in that the extract is concentrated under reduced pressure or lyophilized after extraction.

The present invention also relates to a cosmetic composition, characterized in that the extract is concentrated or lyophilized in 0.001 to 30.0% by weight in at least one solvent selected from purified water, ethanol, butylene glycol and propylene glycol.

In addition, the present invention after adjusting the pH of the culture medium at the time of the main culture to 5.0 ~ 7.0, inoculate the Ganoderma lucidum mycelium so as to 4.0 to 6.0% (v / v), the temperature 22 ~ 32 ℃, rotation speed 120 ~ 180rpm, It relates to a cosmetic composition characterized by culturing for 5 to 7 days in aeration 1.2 ~ 1.8vm conditions.

The present invention also relates to a cosmetic composition for preventing hair loss and promoting hair growth containing the fermented young flower extract.

In addition, the present invention relates to a cosmetic composition exhibiting a skin irritation-releasing effect containing the fermented young flower extract.

In addition, the present invention, the cosmetic composition is in the form of hair or scalp-related products, such as hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair rinse, hair pack and hair Characterized in that the cosmetic composition selected from the treatments.

The cosmetic composition containing the fermented young flower extract of the present invention promotes radical scavenging effect, antimicrobial effect, hair growth and hair root cell activity without irritating the scalp, and in case of hair loss due to inflammatory expression, through anti-inflammatory action It has a function of promoting hair growth.

In addition, the fermented young flower extract of the present invention is excellent in inhibiting skin cell death and skin stimulation effect to reduce skin irritation caused by the cosmetic substrate.

Figure 1 shows the morphological changes of the hair growth process according to the application of the sample obtained in Example 1 compared to the sample untreated control and positive control (3% minoxidil).
Figure 2 is a micrograph showing the histological changes of the hair follicle tissue according to the application of the sample obtained in Example 1 and the positive control group (3% minoxidil) and the sample.

Hereinafter, the present invention will be described in more detail by way of examples. It should be noted, however, that these examples are only illustrative examples of the present invention, and the scope of the present invention is not limited to these examples.

< Comparative Example  1> bottlewood extract manufacturer

After dipping, 500 g of the dried birches were refluxed three times for 3 hours with a hot 95% (V / V) ethanol aqueous solution, and the mixture was cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C. or lower to obtain 78 g of ethanol extract. The flower petal extract was prepared using one or more solvents selected from purified water, ethanol, butylene glycol, and propylene glycol to contain 15.0% by weight of a vacuum concentrate and a freeze-dried product.

< Example  1> Preparation of Fermented Flowering Tree Extract

Ganoderma lucidum mycelium was inclined in a test tube containing potato dextrose agar medium and stored at 4 ℃ and used subcultured every month. Aseptic homogenization of mycelial subcultured in the slope medium, inoculated so that the Ganoderma lucidum mycelium 5% (v / v) in the mixed medium of the locust extract and yeast-wort obtained in Comparative Example 1 and then fermented The culture was carried out for 6 days under conditions of a temperature of 25 ° C., a rotational speed of 150 rpm, aeration rate of 1.5vm, and pH 7.0. After culturing, the Ganoderma lucidum mycelium was removed from the culture broth, and then fermented cauliflower extract was obtained.

< Experimental Example  1> Determination of Components and Contents in Fermented Prunus japonica Extract

In Experimental Example 1, the qualitative analysis of phenolic substances, flavonoids, flavonoid glycosides, etc., and total phenolic contents and total flavonoid contents among the main components of the fern extract prepared in Comparative Example 1 and the fermented vine extract prepared in Example 1 Were compared by quantitative analysis.

First, qualitative analysis of phenolic substances, flavonoids, flavonoid glycosides, etc. was performed as follows.

(1) Qualitative analysis of phenolic substances and flavonoids

When 1 to 2 drops of 2.5% FeCl ₃ ethanol solution was added to the extracts of Comparative Example 1 and Example 1, the color was dark green or the formation of precipitates was observed.

(2) Qualitative Analysis of Flavonoid Glycosides

Using concentrated sulfuric acid method, 5 ml of concentrated sulfuric acid was immersed in the extracts of Comparative Example 1 and Example 1 to observe the formation of yellow and yellow-red precipitates.

(3) Sugar Qualitative Analysis

Two to three drops of 5% α-naphthol alcohol solution was added to the extracts of Comparative Example 1 and Example 1, respectively, and when the concentrated sulfuric acid was added through the walls, it was observed whether reddish purple appeared at the interface.

(4) Determination of Total Phenolic Content

10 ml of distilled water was added to 1 ml of the extracts of Comparative Example 1 and Example 1, 2 ml of Folin-Ciocalteu phenolic formulation was added, mixed, and reacted at room temperature for 5 minutes. 2 ml of 20% sodium carbonate was added to the reaction mixture, followed by reaction at room temperature for 1 hour, and then absorbance at 725 nm was measured. At this time, tannic acid was used as an indicator.

(5) Determination of total flavonoid content

0.5 ml of the extract of Comparative Example 1 and Example 1 were mixed with 0.1 ml of 10% aluminum nitrate, 0.1 ml of 1 M potassium acetate, and 4.3 ml of ethanol, respectively, and reacted at room temperature for 40 minutes, and then absorbance at 415 nm. Measured. At this time, the indicator material was used quercetin (quercetin).

Name of sample Total phenol content
(Μl / ml of extract) Total Flavoid Content
(Μl / ml of extract) Comparative Example 1 719.91 644.87 Example 1 929.00 733.13

As a result of measuring the total phenol content and the total flavonoid content (Table 1), it was confirmed that the total phenol content and total flavonoid content of the extract of Example 1 was increased than the extract of Comparative Example 1.

From the above results, it could be confirmed that the fermented young leaf extract prepared by simple extraction with Ganoderma lucidum mycelium could increase the total phenolic content and the total flavonoid content. Based on this, the cauliflower fermented by Ganoderma lucidum mycelium To prepare a cosmetic composition for preventing hair loss of the present invention, characterized in that it contains an extract.

< Experimental Example  2> DPPH Radical  Antioxidant Effect Measurement Experiment

In this experiment, the scavenging effect on the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical was measured by slightly modifying the method used by Yoshida et al. (1989) to measure the antioxidant effect of the material obtained in Example 1. . Since DPPH is purple as a stable free radical, it is a measure of the extent to which purple is removed as the sample scavenges radicals.

First, DPPH was dissolved in methanol at a concentration of 1 × 10 ⁻⁴ M and 0.15 ml of this solution was placed in a test tube. 0.15 ml of each concentration sample was added thereto, and the absorbance was measured at 565 nm after 20 minutes of reaction at room temperature. Green tea extract was used instead of the material of the example as a positive control. The free radical scavenging effect of each sample was expressed as SC ₅₀ by calculating the concentration of 50% scavenging of radicals using Equation 1. At this time, the negative control was taken as the reaction absorbance of the liquid not treated with the sample.

Name of sample SC ₅₀ (μl / ml) Example 1 35 Green tea extract 400

As shown in Table 2, the fermented young flower extract of the present invention showed a very strong activity of about 10 times as compared to the green tea extract as compared to the green tea extract, which is a powerful antioxidant.

< Experimental Example  3> free radicals Radical  Antioxidant Effect Measurement Experiment

This Experimental Example shows the activity produced by the xanthine-xanthine oxidase system by slightly modifying the method used by Furuno et al. (2002) to measure the antioxidant effect of the material obtained in Example 1. The scavenging effect on the superoxide radical was measured. The free radicals produced by the xanthine and xanthine oxidase reactions react with Nitro Blue Tetrazolium (NBT) to give a blue color, so the amount of blue reduced as the sample eliminates free radicals is measured. It is.

First, 3 × 10 ^-3 M xanthine, 3 × 10 ^-4 M EDTA, 7.5 × 10 ^-4 M NBT (nitroblue tetrazolium), and 0.15 mg / mL bovine serum albumin (BSA) solution were prepared in each concentration sample. The mixture was added and mixed, followed by reaction at room temperature for 10 minutes. Thereafter, xanthine oxidase (0.25 U / ml) solution was added to each test tube and reacted at room temperature for 20 minutes, and then absorbance was measured at 565 nm. Green tea extract was used instead of the material of the example as a positive control. The active oxygen radical scavenging effect of each sample was expressed as SC ₅₀ by calculating the concentration of 50% scavenging radicals using Equation 2. At this time, the negative control was taken as the reaction absorbance of the liquid not treated with the sample.

Name of sample SC ₅₀ (μl / ml) Example 1 20 Green tea extract 340

As shown in Table 3, the fermented young flower extract of the present invention showed a very strong activity of about 16 times as compared to the green tea extract.

From the above results, it was confirmed that the excellent antioxidant effect of the fermented young flower extract of the present invention.

< Experimental Example  4> Antimicrobial effect measurement experiment

In this experimental example, the antimicrobial effect of the samples obtained in Example 1 was evaluated by the minimal inhibitory concentration (MIC) test by slightly modifying the method used by Cho et al. (2001).

In a way that while the concentration of the sample 2-fold serial dilution in order to measure the minimum inhibitory concentration measuring MIC, cause of dandruff and seborrheic dermatitis, M. Les trick other (M. restricta), M. MIC for M. furfur was measured. Prepare the strain suspension by diluting the pre-cultivated test bacteria to the liquid medium to about 1 × 10 ⁶ CFU / ㎖, and then diluting serially by 2 times 100ml each sample in each well of the 96-well U-type microplate In addition, 100 μl of test bacteria were added to each well, and then cultured in a 35 ° C. incubator for 48 hours. The concentration at which no growth of bacteria was observed at 630 nm was determined by MIC of the sample by UV / Vis fluorescence spectrophotometer.

Strain name MIC (%) M. restricta 0.2 M. furfur 0.4

As shown in Table 4 it can be seen that the fermented young flower extract of the present invention has an excellent antibacterial effect.

< Experimental Example  5> 5- Lipooxygenase  (5- LO Inhibitory activity measurement experiment

This experiment was performed by slightly modifying the method used by Block et al. (1988) to measure the anti-inflammatory effects of the samples obtained in Example 1.

First, 20 μl of sample and 20 μl of soy lipooxygenase (Type V, 200 units / final concentration) were added to 1000 μl of reaction buffer (0.1M Tris-HCl, pH8.5), followed by pre-reaction at 25 ° C. for 2 minutes. 30 μl of linoleic acid was added so that the final concentration was 110 μM, and the initial reaction rate was obtained by measuring absorbance at 234 nm at 20 ° C. for 3 minutes at 25 ° C. based on the added time. As a positive control, nordihydroguaiaretic acid (NDGA) was used instead of the material of the example. The anti-inflammatory effect of each sample was expressed as the inhibition rate of 5-LO using Equation 3. At this time, the negative control was taken as the reaction absorbance of the liquid not treated with the sample.

Sample name (µl / mL) 5-LO inhibition rate (%) Example 1 500 82.5 250 76.9 NDGA 500 98 250 90

As shown in Table 5, the fermented young flower extract of the present invention showed a very good inhibitory activity compared to NDGA, a potent 5-LO inhibitor.

< Experimental Example  6> Hair growth promoting effect measurement experiment

This experimental example measured the hair growth promoting effect of the sample obtained in Example 1 and a positive control containing 3% minoxidil (minoxidil) which is generally used as a hair loss prevention and hair growth promoter.

First, 6-week-old black mice (C57BL / 6, male), which are dormant hairs having a dorsal skin color of pink, were purified for 7 days, and then the hairs of the dorsal area were shaved, and 6 rats were assigned to each experimental group. The next day, 0.2 ml each of the sample and the positive control obtained in Example 1 were sufficiently applied using a brush in the central part such as hair cutting. After each sample was applied for about 3 minutes to drink and rinsed thoroughly with distilled water. Sample treatment was conducted twice daily and the condition was observed for 3 weeks. Since the efficacy can not be verified when the hair growth progresses anymore, the experiment was performed for about 3 weeks, and the experiment was terminated when the hair growth of the sample having the highest efficacy progressed more than 95%. To compare hair length and hair growth over time, negative controls were observed in their natural growth without application of samples.

Name of sample Indicators Elapsed days 1 parking 2 parking 3 parking Example 1 Hair length (mm) 1.11 ± 0.13 2.49 ± 0.17 3.95 ± 0.29 Growth rate (%) 27.1 60.7 96.3 Positive control (3% minoxidil) Hair length (mm) 1.38 ± 0.16 2.91 ± 0.32 3.73 ± 0.40 Growth rate (%) 33.6 71.0 91.0 Negative control group Hair length (mm) 0.55 ± 0.15 1.30 ± 0.25 2.13 ± 0.34 Growth rate (%) 13.4 31.7 51.9

As shown in Table 6 and Figure 1, the fermented young flower extract of the present invention showed an excellent hair growth effect than minoxidil used as a hair growth promoter.

< Experimental Example  7> Experiment to measure hair follicle cell activation

In Experimental Example 6, three rats were examined at the end of the seventh day of the test and three rats were examined at the end of the test (21 days) while the hair growth promoting experiment was conducted in Experimental Example 6, and the degree of hair root cell activity promotion of each sample was measured.

After extracting the sample application site by using an anatomizer and fixing with formalin, dehydrating with alcohol and xylene step by step to remove paraffin, and then made a 5 μm section using a tissue sectioner. Again paraffin was removed with alcohol and xylene. The tissues were stained with H & E (Hematoxylin & Eosin) to observe histological changes of hair follicle tissues under an optical microscope. The results of measuring the degree of promoting hair follicle activity are shown in FIG. 2. According to Figure 2, the fermented young flower extract of the present invention was found to be superior to the hair root cell activity promoting effect than minoxidil used as a hair growth promoter.

< Experimental Example  8> Skin irritation test

In order to measure the skin irritation of the sample obtained in Example 1, this experimental example was determined using toxicological test criteria such as medicines, and Draize's PII (Primary Irritation Index) of Draize generally used the degree of irritation to the skin. Evaluated by calculation method (0; no stimulation, 5; strong stimulation).

Rabbits were depilated 24 hours before each sample was treated, and divided into treatment and control sections of normal skin and wound skin. The coating method was applied once to the application site by 1g of sample per rabbit. The control group was applied with the same amount of sterile saline and covered with gauze. In order to suppress evaporation of the sample, it was covered with a solid material foil having no permeability and reactivity, and fixed with a tape and then applied for 24 hours, 72 hours, and 120 hours.

Name of sample PII result 24 hours 48 hours 120 hours Example 1 0.3 (no irritation) 0.1 (no irritation) 0.1 (no irritation) Control group 0.1 (no irritation) 0.1 (no irritation) 0.1 (no irritation)

As shown in Table 7, the fermented vine extract of the present invention exhibited hypoallergenicity of 0.3 with PII of 0.3 for 24 hours, and 0.1 stimulation of 0.1 for 120 hours of application, and no irritation of 0.1 for 120 hours. The extract was confirmed to have less irritation to the skin.

Claims (7)

Byeongkkot trees (Weigela subsessilis ) extracting the extract;
Ganoderma lucidum ( Ganoderma) in the extract lucidum ) Mycelium or Ganoderma lucidum ( Ganoderma) lucidum ) mixing the whole culture solution of the mycelium, and then main culture; And
After the main culture, Ganoderma lucidum ( Ganoderma) from the main culture lucidum ) removing the mycelia; hair loss preventing and hair growth cosmetic composition comprising a filtrate obtained from the active ingredient.
The method according to claim 1,
Hair extraction prevention and hair growth promoting cosmetic composition, characterized in that using an aqueous solution containing 40 ~ 95% (v / v) ethanol as the extraction solvent during the extraction.
The method according to claim 1,
Hair loss prevention and hair growth cosmetic composition, characterized in that the content of the filtrate is 0.001 ~ 30.0% by weight based on the entire cosmetic composition.
The method according to claim 1,
The extract is hair loss prevention and hair growth cosmetic composition, characterized in that concentrated under reduced pressure or lyophilized after extraction.
The method of claim 4,
The reduced pressure concentrated or lyophilized extract is a cosmetic composition for preventing hair loss and promoting hair growth, characterized in that dissolved 0.001 ~ 30.0% by weight in any one or more solvents selected from purified water, ethanol, butylene glycol and propylene glycol.
The method according to claim 1,
The main culturing step is to adjust the pH of the culture medium to 5.0 ~ 7.0, Ganoderma lucidum ( Ganoderma lucidum ) After inoculating mycelium 4-6% (v / v), hair loss prevention, characterized in that incubated for 5-7 days at a temperature of 22 ~ 32 ℃, rotation speed 120 ~ 180rpm, aeration 1.2 ~ 1.8vm conditions And cosmetic composition for promoting hair growth.
The method according to claim 1,
The cosmetic composition is hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair rinse, hair pack and hair treatment cosmetics for hair loss prevention, characterized in that selected from Composition.

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